Comparison of growth rates of bovine retinal and brain microvascular pericytes in different oxygen concentrations in vitro.

Abstract

The hyperoxic injury of the microcirculation in the central nervous system appears to be specific to the retina in premature mammals. Oxygen tensions in normal adult mammalian retina and brain vary between nearly 0 and 90 mmHg. This study sought to compare the in vitro replication of retinal and brain microvascular pericytes in normal glucose medium and in 1%, 5% and 20% oxygen (equivalent to 15 mmHg, 35 mmHg and 150 mmHg, respectively). A preliminary study, using oxygen microelectrodes, confirmed that the pericellular oxygen tension of pericytes, cultured in medium under air, was within 13 mmHg of the tension of the gas phase above the media. Pericytes were highly enriched by magnetic antibody cell sorting with the anti-pericyte monoclonal antibody (3G5) to 95% to 99% purity, to remove cell contaminants which may have invalidated the mitogenic assay. Mitogenic assays showed that brain pericytes replicated faster than their counterparts from retina (P < 0.0001, averaged for data from all culture conditions using three-way ANOVA). Reduction of oxygen tension from 150 to 15 mmHg led to significantly increased replication of retinal pericytes (P = 0.01), but an insignificant increase for brain pericytes. We have found that pericytes from the brain and retina cultured conventionally in fetal calf serum consume a relatively low amount of oxygen. Decreasing the oxygen tension to 1% (15 to 20 mmHg) increased the replication of retinal pericytes but not brain pericytes in normal glucose concentrations and in fetal calf serum. That retinal pericyte replication is sensitive to variation in oxygen tensions, indicates that the retinal microvascular cells have a unique biological response. This growth sensitivity to oxygen may be important in the pathogenesis of retinopathy of prematurity.